Journal of Zhejiang University SCIENCE
(ISSN 1009-3095, Monthly)

2005   Vol. 6B   No. 9   p.859-864

            [ Home Page ] | [ PDF Full Text ]   On-line Access Date:   Aug. 31, 2005

Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2

JIANG Qiang†, ZHANG Su-zhan†‡, PENG Jia-ping, WANG Xu-lin

(Cancer Institute, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China)
 †E-mail: jiangtiger@zju.edu.cn; zhangscy@tom.com
Received Apr. 28, 2005; revision accepted May 24, 2005

Abstract: Objective: To prepare microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2 (TIMP-2), and investigate their biological characteristics in vitro. Methods: Chinese hamster ovary (CHO) cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) assay. Enzyme linked immunosorbent assay (ELISA) and reverse zymography were used to confirm the release of biologically active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl sulfoxide (DMSO) as preservative agent. Results: The microcapsules appeared like a sphere with diameter of 300~600 μm. The surface of the capsule wall was clearly smooth. The microencapsulated cells survived well and kept proliferating over the 6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse zymography confirmed the bioactivity of MMP (matrix metalloproteinase) inhibition of TIMP-2. The cryopreservation process did not damage the microcapsule morphology nor the viability of the cells inside. Conclusion: Microencapsulated engineered CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules.

Key words: Microencapsulation, Recombinant cells, Human tissue inhibitor of metalloproteinase-2, Cell culture
doi:10.1631/jzus.2005.B0859             CLC number: R73


‡ Corresponding Author